Quantitative analysis of fibroblast morphology on microgrooved surfaces with various groove and ridge dimensions

Abstract

Fibroblasts have been shown to respond to substratum surface roughness. The change in cell size, shape and orientation of rat dermal fibroblasts (RDF) was therefore studied using smooth and microtextured silicone rubber substrata. The microtextured substrata possessed parallel surface microgrooves that ranged in width from 1.0 to 10.0/im, and were separated by ridges of 1.0 to 10.0¿¿m. The grooves were either 0.45 or 1.00^m deep. Prior to incubation, the substrata were cleaned and given a radio frequency glow discharge treatment. After surface evaluation with scanning electron microscopy and confocal laser scanning microscopy, RDF were incubated on these substrata for 5 days. During this period of incubation, the RDF were photographed on days 1, 2, 3, 4, and 5, using phase contrast microscopy. Digital image analysis of these images revealed that on surfaces with a ridge width < 4 .0 ¿tm, cells were highly orientated (<10") and elongated along the surface grooves. H Protrusions contacting the ridges specifically could be seen. If the ridge width was larger than 4.0/tm, cellular orientation was random («45") and the shape of the RDF became more circular. Furthermore, results showed that the ridge width is the most important parameter, since varying the groove width and groove depth did not affect the RDF size, shape, nor the angle of cellular orientation. ((■) 1996 Elsevier Science Limited

Interactive effects of surface topography and pulsatile electrical field stimulation on orientation and elongation of fibroblasts and cardiomyocytes

“…In addition, ECM proteins orient parallel to the groove direction on microstructured surfaces, affecting the orientation of integrins. Since adhesion molecules are coupled to the actin cytoskeleton, this causes the orientation of the actin filaments parallel to the groove direction [12,13], as we also observed in Figure 5. Numerous previous studies demonstrated that polymerization of F-actin is critical in cell motility and response of fibroblasts to surface topography [12,55,56].…”

“…focal adhesions) to an orientation parallel to the groove direction [12,13]. As the ridge size of the grooved surfaces is decreases (to e.g.…”

“…Microstructured grooves were used previously in 2D to direct orientation and elongation of fibroblasts [10][11][12][13]. The process was dependent upon the orientation of actin cytoskeleton [12] that also oriented along the direction of the grooves.…”

A new approach to study fibroblast migration

“…First, the current technique may be combined with time-lapse video-microscopy so that cell migration can be continuously monitored. In addition, because microgroove dimension (groove width and depth) may affect cell orientation and motility [den Braber et al, 1996;Dalton et al, 2001;Yoshinari et al, 2003], the modification of current microgroove dimensions for studying motility of different types of cells may be needed. Finally, since TGF-b1 and TGF-b3 induce differential contraction in tendon fibroblasts [Campbell et al, 2004], their possible differential effects on fibroblast motility can be investigated using the current technique.…”

Competitive Guidance Cues Affect Fibroblast Cell Alignment: Electric Fields vs. Contact Guidance

“…In general, cells become highly oriented along grooves with widths between 1 and 10 gim. There is some disagreement, however, as to whether groove depth is the most significant factor in guiding cell alignment [3,4]. Another physiologically relevant guidance cue is an electric field (EF), which is present in many tissues during development and wound repair [5].…”